1. Field of the Invention
This invention relates to a new method for the quantitative determination of polyamines. More particularly, it relates to a method which can accurately determine the amount of polyamines by a simple process.
2. Description of the Prior Art
Polyamines, widely distributed in living bodies, are substances of medical importance because these are found in large quantities in proliferating cells, especially in tumor cells. Russell, et.al. reported in 1975 that the body fluids (e.g., urine and blood) of cancer patients, as compared with normal persons, contain a larger amount of polyamines. Since then, many researchers have studied the correlation between cancer and the content of polyamines in body fluids, and confirmed the validity of the report by Russell, et.al. Quantitative determination of polyamines in body fluids is very difficult because of the extremely low concentration of polyamines and the presence of many other substances in the test sample. Recently, an enzymatic method has received attention as a means to rapidly determine the amount of polyamines in body fluids. In a typical example, a sample solution containing polyamines is allowed to react with an enzyme capable of oxidizing polyamines (hereinafter referred to as polyamine oxidizing enzyme) to form hydrogen peroxide, which is then introduced to a color-developing system comprising 4-aminoantipyrine, phenol and a peroxidase, followed by colorimetric measurement of the dye formed.
The problem involved in this method is that the hydrogen peroxide formed tends to undergo decomposition by reducing agents present in body fluids, such as ascorbic acid and uric acid. Hence, these reducing agents must be previously removed or separated from the sample solution to obtain an accurate determination of polyamines, thus complicating the operations involved and increasing the time required for analysis. In addition, this method, based on the measurement of hydrogen peroxide, has an inherent problem resulting in measurement errors due to the unavoidable lost of this unstable compound.